LAMP detection primers, system and method for detecting health state of exopalaemon carinicauda after WSSV (White Spot Syndrome Virus) infection

A technology of white shrimp, health status, applied in biochemical equipment and methods, recombinant DNA technology, microbial determination/inspection, etc., can solve the problems of fluctuation, increase, decrease, etc. low cost effect

Pending Publication Date: 2017-11-21
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

These parameters are often in a relatively stable range in the healthy state of the human body, but when the body receives pathogen invasion or other external adverse stimuli, it will lead to an increase or decrease of one or some of the above indicators. abnormal fluctuation

Method used

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  • LAMP detection primers, system and method for detecting health state of exopalaemon carinicauda after WSSV (White Spot Syndrome Virus) infection
  • LAMP detection primers, system and method for detecting health state of exopalaemon carinicauda after WSSV (White Spot Syndrome Virus) infection
  • LAMP detection primers, system and method for detecting health state of exopalaemon carinicauda after WSSV (White Spot Syndrome Virus) infection

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Experimental program
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Effect test

Embodiment 1

[0041] The screening of the genes related to the health status in the white shrimp and the construction process of the LAMP detection system based on the genes are as follows:

[0042] (1) According to the transcriptome data of white spot syndrome virus (White Spot Syndrome Virus, WSSV) before and after infection of white spot syndrome virus (WSSV), 57 sequences with significant differences in the expression level of transcript levels before and after infection were selected as candidate genes, and primers were designed respectively. The samples before and after WSSV infection were detected by real-time fluorescent quantitative PCR.

[0043](2) According to the results of the preliminary screening, seven genes whose transcription levels changed significantly after infection with pathogens were selected for statistical detection and verification of a single sample. Select 100 cultured healthy white prawns (average body weight, 2.8±0.4g) and raise them temporarily for 3 days. Af...

specific Embodiment 2

[0053] (1) Take approximately 0.2 g gill filaments of shrimps S1 and S2 to be tested and healthy shrimps S3 and S4 respectively, place them in sampling tubes, and grind them quickly with a grinding rod to a slurry state;

[0054] (2) Pick the diaphragm with a toothpick, place it in the sample slurry, and after it is fully wetted, pick it back into the diaphragm tube with a toothpick;

[0055] (3) Absorb absolute ethanol with a straw, drop 2 to 3 drops on the wet sampling diaphragm in the tube, flick the bottom of the tube to stir the diaphragm for 30 seconds;

[0056] (4) Use a new toothpick to transfer the above-mentioned sampling membrane to the rinse tube with the corresponding number, and shake the rinse tube for 3 to 4 minutes;

[0057] (5) Use a new toothpick to transfer the membranes in the above-mentioned rinsing tubes, the positive control FTA membranes in the positive and negative control tubes, and the negative control FTA membranes to the corresponding numbered nuc...

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Abstract

The invention relates to an LAMP detection method for detecting the health state of exopalaemon carinicauda before and after white spot syndrome virus infection. According to the LAMP detection method, in allusion to a cDNA sequence of a gene of which the expression quantity is significantly increased in the early stage of WSSV outburst and which is obtained through massive screening in the exopalaemon carinicauda, 6 primers are designed, and a (Loop-mediated isothermal amplification, LAMP) detection method is established. By the LAMP detection method, nucleic acid extraction and purification are not required, the cost is low, the operation is convenient, and the detection process is fast and sensitive. By the LAMP detection method, diagnosing and prewarning effects can be quickly and effectively achieved in the early stage of the disease outburst of the WSSV-infected exopalaemon carinicauda.

Description

technical field [0001] The invention relates to a LAMP detection method for assessing the health status of white prawns infected with white spot syndrome virus. Background technique [0002] White prawn is an important small-scale economic shrimp in the coast of mainland my country. It has the characteristics of fast growth and delicious taste. However, with the expansion of breeding scale, disease problems have become a serious obstacle to industrial development. White spot syndrome virus (WSSV) is one of the most serious pathogens. WSSV belongs to the genus Whispovirus of the Nimaviridae family. It is a circular double-stranded DNA virus that can infect various shrimps such as white prawns, prawns, and crayfish. Once infected with WSSV, the host's mortality rate will reach 100% within 7-10 days after being infected with WSSV, which will bring serious economic losses to the shrimp farming industry. The current diagnostic methods for WSSV mainly use PCR, realtime-PCR and o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6844C12Q1/6883C12Q2600/158C12Q2531/119
Inventor 李富花刘飞李诗豪
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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